Rocha A, Ruiz S, Coll J M
INIA, SGIT, Dept. Biotecnología, Madrid, Spain.
J Physiol Biochem. 2002 Mar;58(1):45-56. doi: 10.1007/BF03179837.
The increasing use of cationic liposomes as vectors for DNA transfection of eukaryotic cells is due to its high efficiency and reproducibility. After the interaction of the DNA cationic-liposome complexes (DNA-CLC) with the plasma membrane, the entry into the cells delivers the DNA-CLC to the endosome-lysosome pathway where some of the DNA-CLC are degraded. The non-degraded DNA that escapes to the cytoplasm, still has to transverse the nuclear membrane to be transcribed and then translated. To improve the efficiency of the whole process, we can manipulate the DNA (sequences, promoters, enhancers, nuclear localisation signals, etc), the DNA-CLC (lipids) or the plasmatic, endosomal and/or nuclear cellular membranes (ultrasound, electroporation, Ca++, pH of the endosomes, mitosis, fusogenic peptides, nuclear localisation signals, etc). Most of these methods have been generally used individually but in combination, may greatly improve the efficiency and reproducibility of in vitro transfection. While much of this work remains yet to be done and present results further explored, the application of these efforts is essential to the future development of new gene therapy strategies.
阳离子脂质体作为真核细胞DNA转染载体的使用日益增加，这归因于其高效率和可重复性。DNA阳离子脂质体复合物（DNA-CLC）与质膜相互作用后，进入细胞将DNA-CLC传递至内体-溶酶体途径，其中一些DNA-CLC会被降解。逃脱至细胞质的未降解DNA，仍需穿过核膜才能被转录进而翻译。为提高整个过程的效率，我们可以操控DNA（序列、启动子、增强子、核定位信号等）、DNA-CLC（脂质）或细胞质、内体和/或细胞核细胞膜（超声、电穿孔、钙离子、内体pH值、有丝分裂、融合肽、核定位信号等）。这些方法大多通常单独使用，但联合使用可能会大大提高体外转染的效率和可重复性。虽然这项工作的大部分仍有待完成且现有结果需进一步探索，但这些努力的应用对于新基因治疗策略的未来发展至关重要。
